CRISPR Custom Library

Tailor-Made to Precisely Match Your Unique Research Needs

When your research involves rare disease genes, non-model organisms, specific mutation sites, or multiple CRISPR modalities, standard libraries fall short. GeneRulor's CRISPR Custom Library Service offers fully personalized solutions tailored to your exact needs — from gene selection and sgRNA design to vector construction.

Figure 1 CRISPR Custom Library Service Workflow and Core Capabilities

1. What is a CRISPR Custom Library?

A CRISPR Custom Library is an sgRNA library built from scratch for specific research needs — whether targeting a defined gene set, mutation sites, non-model organisms, or multiple CRISPR modalities.

2. Core Advantages of Custom Libraries

2.1 Fully Personalized Design

• Customizable sgRNA count per gene

• Freely select target genes: From dozens to thousands

• Site-specific targeting: Exons, domains, or mutation hotspots

• Multi-modality integration: KO, CRISPRi, CRISPRa in one library

• Species flexibility: Any sequenced species

2.2 Professional Design Algorithms

• Latest sgRNA algorithms: Activity prediction, off-target assessment, chromatin accessibility

• Optimized for your screen type (survival, proliferation, phenotype, etc.)

• Challenging gene handling: Abnormal GC content, repeats, etc.

• Species-specific PAM and genomic feature optimization

2.3 Flexible Vector Selection

• Multiple vector backbones: lentiCRISPR, lentiGuide, and more

• Diverse promoters: Constitutive, inducible, tissue-specific

• Customizable markers: Antibiotic resistance, fluorescent proteins, etc.

• Special modules: Barcodes, reporter genes, bidirectional promoters, etc.

2.4 Stringent Quality Control

• Design review by postdoctoral team

• Synthesis verification by Sanger sequencing

• Library NGS validation for coverage and uniformity

2.5 End-to-End Technical Support

• Design phase: Objective discussion and design recommendations

• Construction phase: Real-time updates and issue reporting

• Validation phase: Quality reports and usage recommendations

• Application phase: Protocol optimization and data analysis

3. GeneRulor Custom Library Service Scope

3.1 Targeted Gene Set Libraries

Tailored to your research question:

• Candidate gene validation libraries: From prior screens, GWAS studies, or literature mining

• Pathway-specific libraries: Specific pathways or processes (e.g., autophagy, DNA repair)

• Disease-associated libraries: All known causative genes for a specific disease

• Chromosomal region libraries: All genes in a chromosomal region for fine-mapping

3.2 Non-Model Organism Libraries

For any sequenced species of interest:

• Agricultural species: Rice, wheat, tomato, pig, cattle, etc.

• Aquatic organisms: Zebrafish, shrimp, fish, etc.

• Microorganisms: Bacteria, fungi, yeast, etc.

• Emerging model organisms: Primates, amphibians, etc.

3.3 Special Site-Targeting Libraries

Precision targeting to specific sequences:

• Mutation site libraries: Targeting oncogenic mutations (KRAS G12D, TP53 hotspots, etc.)

• Domain tiling libraries: High-density tiling across protein functional domains

• Exon-targeting libraries: Knockout of specific exons or splice sites

• Non-coding region libraries: lncRNAs, enhancers, promoters, and other regulatory elements

3.4 Multi-Modality Integrated Libraries

Multiple modalities in one library:

• KO + CRISPRi: Parallel comparison of permanent knockout and reversible knockdown in the same gene set

• CRISPRi + CRISPRa: Simultaneously investigate the effects of gene silencing and activation

• Base Editor libraries: C-to-T or A-to-G base editor libraries for saturation mutagenesis screens

• Prime Editor libraries: Precise insertion, deletion, or replacement of specific sequences

3.5 Combinatorial Screening Libraries

For studying gene-gene interactions:

• Dual-sgRNA libraries: Two sgRNAs per vector for pairwise interaction screens

• Synthetic lethality screens: Identify synthetic lethal partners in a mutant background

• Suppressor/enhancer screens: Identify genetic modifiers of a given phenotype

4. GeneRulor One-Stop Custom Service Workflow

4.1 Phase 1: Needs Analysis and Solution Design (1–2 Weeks)

4.1.1 In-Depth Needs Assessment

• Understand background, objectives, and expected outcomes

• Clarify target genes, species, and cell lines

• Discuss special requirements and technical challenges

4.1.2 Professional Solution Design

• Evaluate project feasibility

• Provide multiple design options

• Explain rationale and expected outcomes

• Provide quotation and timeline

4.1.3 Solution Confirmation

• Adjust based on feedback

• Confirm all design details

• Sign technical and NDA agreements

4.2 Phase 2: Library Construction and Validation (8–12 Weeks)

4.2.1 sgRNA Design (1–2 Weeks)

• Generate candidate sgRNAs

• Off-target analysis and activity prediction

• Manual review and optimization

4.2.2 Library Synthesis (4–6 Weeks)

• High-throughput oligo synthesis

• Vector cloning and amplification

• Quality control testing

4.2.3 NGS Validation (2–3 Weeks)

• Deep sequencing (typically >1000X coverage)

• Coverage and uniformity analysis

• Detailed validation report

4.2.4 Viral Packaging (Optional, 1–2 Weeks)

• High-titer lentivirus preparation

• Titer testing and QC

• Aliquoting and cryopreservation

4.3 Phase 3: Delivery and Technical Support

4.3.1 Complete Delivery

• Library plasmid or virus

• Design documentation and sgRNA list

• NGS validation report

• Recommended experimental protocol

4.3.2 Ongoing Technical Support

• Experimental operation guidance

• Problem diagnosis and resolution

• Data analysis assistance

• Follow-up optimization recommendations

5. Custom Libraries vs. Off-the-Shelf Libraries

How to Decide?

Choose a Custom Library if you:

• Studying non-model organisms or special species

• Need a library for a specific gene set or mutation sites

• Need to integrate multiple CRISPR modalities

• Special sgRNA design or vector system needs

• Ultra-high-density or tiling design required

• Pioneering project where standard libraries fall short

Choose an Off-the-Shelf Library if you:

• Need rapid project launch

• Conventional model organisms (human, mouse)

• Whole-genome or major functional gene set screens

• Prefer proven libraries to minimize risk

• Limited budget, prioritizing cost-effectiveness

Our team can help evaluate your needs and recommend the optimal solution. In many cases, combining an off-the-shelf library with a few custom sgRNAs offers both speed and flexibility.

6. Why Choose GeneRulor Custom Libraries?

6.1 Expert Team

• Postdoctoral station with strong R&D capacity

• Rich non-model organism library design experience

• Deep understanding of diverse research needs

6.2 Technology Leadership

• Proprietary sgRNA design algorithms

• Continuously updated design and QC standards

• Supports latest CRISPR technologies (base editing, prime editing, etc.)

6.3 Flexible Customization

• Designed entirely to your specifications

• No off-the-shelf constraints

• Accommodates all special requirements

6.4 Quality Assurance

• Rigorous NGS validation

• Optional functional pre-validation

• Detailed quality reports

6.5 Full-Process Support

• Full-cycle service: design to application

• Prompt response and issue resolution

• Optional data analysis service

6.6 Success Stories

• Served over 100 custom projects

• Covering 20+ species

• Supported publications in Nature, Science, and top-tier journals

References

[1] Doench, J. G., et al. (2016). Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nature Biotechnology, 34(2), 184-191.

[2] Sanson, K. R., et al. (2018). Optimized libraries for CRISPR-Cas9 genetic screens with multiple modalities. Nature Communications, 9(1), 5416.

[3] Broad Institute GPP sgRNA Designer: https://portals.broadinstitute.org/gpp/public/analysis-tools/sgrna-design

[4] Hanna, R. E., & Doench, J. G. (2020). Design and analysis of CRISPR-Cas experiments. Nature Biotechnology, 38(7), 813-823.